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Title: Purification and crystallization of the ABC-type transport substrate-binding protein OppA from Thermoanaerobacter tengcongensis

Abstract

Highlights: Black-Right-Pointing-Pointer We truncated the signal peptide of OppA{sub TTE0054} to make it express in Escherichia coli as a soluble protein. Black-Right-Pointing-Pointer Crystals of OppA{sub TTE0054} were grown by sitting-drop vapor diffusion method. Black-Right-Pointing-Pointer The crystal of OppA{sub TTE0054} diffracted to 2.25 A. -- Abstract: Di- and oligopeptide- binding protein OppAs play important roles in solute and nutrient uptake, sporulation, biofilm formation, cell wall muropeptides recycling, peptide-dependent quorum-sensing responses, adherence to host cells, and a variety of other biological processes. Soluble OppA from Thermoanaerobacter tengcongensis was expressed in Escherichia coli. The protein was found to be >95% pure with SDS-PAGE after a series of purification steps and the purity was further verified by mass spectrometry. The protein was crystallized using the sitting-drop vapour-diffusion method with PEG 400 as the precipitant. Crystal diffraction extended to 2.25 A. The crystal belonged to space group C222{sub 1}, with unit-cell parameters of a = 69.395, b = 199.572, c = 131.673 A, and {alpha} = {beta} = {gamma} = 90 Degree-Sign .

Authors:
;  [1];  [2]; ;  [3];  [1];  [1];  [3]
  1. State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Tsinghua University, Beijing 100005, People's Republic of China (China)
  2. (China)
  3. Tsinghua-Peking Joint Center for Life Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, People's Republic of China (China)
Publication Date:
OSTI Identifier:
22207902
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 423; Journal Issue: 1; Other Information: Copyright (c) 2012 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CELL WALL; CRYSTALLIZATION; ESCHERICHIA COLI; IMPURITIES; MASS SPECTROSCOPY; NUTRIENTS; PEPTIDES; PURIFICATION; SYNCHROTRON RADIATION; VAPORS

Citation Formats

Gao, Jinlan, Li, Xiaolu, Tsinghua-Peking Joint Center for Life Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, People's Republic of China, Feng, Yue, Zhang, Bo, Miao, Shiying, Wang, Linfang, E-mail: lfwangz@yahoo.com, and Wang, Na, E-mail: nawang@tsinghua.edu.cn. Purification and crystallization of the ABC-type transport substrate-binding protein OppA from Thermoanaerobacter tengcongensis. United States: N. p., 2012. Web. doi:10.1016/J.BBRC.2012.05.067.
Gao, Jinlan, Li, Xiaolu, Tsinghua-Peking Joint Center for Life Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, People's Republic of China, Feng, Yue, Zhang, Bo, Miao, Shiying, Wang, Linfang, E-mail: lfwangz@yahoo.com, & Wang, Na, E-mail: nawang@tsinghua.edu.cn. Purification and crystallization of the ABC-type transport substrate-binding protein OppA from Thermoanaerobacter tengcongensis. United States. doi:10.1016/J.BBRC.2012.05.067.
Gao, Jinlan, Li, Xiaolu, Tsinghua-Peking Joint Center for Life Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, People's Republic of China, Feng, Yue, Zhang, Bo, Miao, Shiying, Wang, Linfang, E-mail: lfwangz@yahoo.com, and Wang, Na, E-mail: nawang@tsinghua.edu.cn. Fri . "Purification and crystallization of the ABC-type transport substrate-binding protein OppA from Thermoanaerobacter tengcongensis". United States. doi:10.1016/J.BBRC.2012.05.067.
@article{osti_22207902,
title = {Purification and crystallization of the ABC-type transport substrate-binding protein OppA from Thermoanaerobacter tengcongensis},
author = {Gao, Jinlan and Li, Xiaolu and Tsinghua-Peking Joint Center for Life Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, People's Republic of China and Feng, Yue and Zhang, Bo and Miao, Shiying and Wang, Linfang, E-mail: lfwangz@yahoo.com and Wang, Na, E-mail: nawang@tsinghua.edu.cn},
abstractNote = {Highlights: Black-Right-Pointing-Pointer We truncated the signal peptide of OppA{sub TTE0054} to make it express in Escherichia coli as a soluble protein. Black-Right-Pointing-Pointer Crystals of OppA{sub TTE0054} were grown by sitting-drop vapor diffusion method. Black-Right-Pointing-Pointer The crystal of OppA{sub TTE0054} diffracted to 2.25 A. -- Abstract: Di- and oligopeptide- binding protein OppAs play important roles in solute and nutrient uptake, sporulation, biofilm formation, cell wall muropeptides recycling, peptide-dependent quorum-sensing responses, adherence to host cells, and a variety of other biological processes. Soluble OppA from Thermoanaerobacter tengcongensis was expressed in Escherichia coli. The protein was found to be >95% pure with SDS-PAGE after a series of purification steps and the purity was further verified by mass spectrometry. The protein was crystallized using the sitting-drop vapour-diffusion method with PEG 400 as the precipitant. Crystal diffraction extended to 2.25 A. The crystal belonged to space group C222{sub 1}, with unit-cell parameters of a = 69.395, b = 199.572, c = 131.673 A, and {alpha} = {beta} = {gamma} = 90 Degree-Sign .},
doi = {10.1016/J.BBRC.2012.05.067},
journal = {Biochemical and Biophysical Research Communications},
number = 1,
volume = 423,
place = {United States},
year = {Fri Jun 22 00:00:00 EDT 2012},
month = {Fri Jun 22 00:00:00 EDT 2012}
}